Top-down label-free LC-MALDI analysis of the peptidome during neural progenitor cell differentiation reveals complexity in cytoskeletal protein dynamics and identifies progenitor cell markers

在神经祖细胞分化过程中,对肽组进行自上而下的无标记 LC-MALDI 分析,揭示了细胞骨架蛋白动力学的复杂性并识别了祖细胞标记物

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作者:Daniel J Maltman, Sven Brand, Eckhard Belau, Rainer Paape, Detlev Suckau, Stefan A Przyborski

Abstract

In the field of stem cell research, there is a strong requirement for the discovery of new biomarkers that more accurately define stem and progenitor cell populations, as well as their differentiated derivatives. The very-low-molecular-weight (<5 kDa) proteome/peptidome remains a poorly investigated but potentially rich source of cellular biomarkers. Here we describe a label-free LC-MALDI-TOF/TOF quantification approach to screen the very-low-molecular-weight proteome, i.e. the peptidome, of neural progenitor cells and derivative populations to identify potential neural stem/progenitor cell biomarkers. Twelve different proteins were identified on the basis of MS/MS analysis of peptides, which displayed differential abundance between undifferentiated and differentiated cultures. These proteins included major cytoskeletal components such as nestin, vimentin, and glial fibrillary acidic protein, which are all associated with neural development. Other cytoskeletal proteins identified were dihydropyrimidinase-related protein 2, prothymosin (thymosin α-1), and thymosin β-10. These findings highlight novel stem cell/progenitor cell marker candidates and demonstrate proteomic complexity, which underlies the limitations of major intermediate filament proteins long established as neural markers.

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